The invention relates to a method for reconstructing the distribution of fluorophores in a non-homogeneous diffusing object by optical tomography, comprising                illumination of the object with continuous light by a light source,        measurement of the light intensity received by detectors at a fluorescence wavelength and        resolution, at each point of a mesh of a volume at least partially containing the object to be observed, of an equation expressing the relation between the measured light intensity, a parameter representative of the properties of the fluorophores, and transfer functions representative of the energy transfer between each of the sources and the fluorophores and between the fluorophores and detectors.        